Cell culture model for acquired chemoresistance of chronic myelogenous leukemia and related methods for identifying agents to overcome resistance

Abstract

A method of generating a chronic myelogenic leukemia (CML) acquired chemoresistant culture model is provided. Such a method may comprise providing a naïve blast crisis CML cell line; administering/contacting the cell line with a mutation-inducing dose of imatinib; maintaining a culture of the treated cell line for a period of time until the treated cell line relapses and repopulates the culture; and determining the repopulated cell culture is a CML acquired chemoresistant cell line by detecting a BCR-ABL mutation, wherein the acquired chemoresistance is achieved by a BCR-ABL mutation.

What is claimed is:  
     
       1. A culture model of chronic myelogenic leukemia (CML) acquired chemoresistant cells, the model generated by:
 contacting a blast crisis CML cell line, which has previously not been exposed to imatinab before, with a dose of imatinib sufficient to induce a BCR-ABL mutation, wherein the blast crisis CML cell line is a KCL-22 cell line; 
 maintaining a culture of the imatinib-treated cell line for about two to three weeks until the cell line relapses and repopulates the culture; and 
 determining that the repopulated cell culture is a CML acquired chemoresistant cell line by detecting a BCR-ABL mutation. 
 
     
     
       2. The model of  claim 1 , wherein the mutation-inducing dose of imatinib is about 2.5 μM or higher. 
     
     
       3. The model of  claim 1 , wherein the mutation-inducing dose of imatinib is between about 2.5 μM and about 10 μM. 
     
     
       4. The model of  claim 1 , wherein the mutation-inducing dose of imatinib is selected from about 2.5 μM, about 5 μM and about 10 μM. 
     
     
       5. The model of  claim 1 , wherein the BCR-ABL mutation is a BCR-ABL kinase domain mutation. 
     
     
       6. The model of  claim 5 , wherein the BCR-ABL kinase domain mutation is a T315I BCR-ABL mutation. 
     
     
       7. The model of  claim 6 , wherein the BCR-ABL kinase domain mutation is a E255K or a Y253H BCR-ABL mutation or a combination thereof. 
     
     
       8. The model of  claim 1 , wherein the BCR-ABL kinase domain mutation is a E255K or a Y253H BCR-ABL mutation or a combination thereof. 
     
     
       9. The model of  claim 1 , wherein the BCR-ABL mutation is stable. 
     
     
       10. The model of  claim 1 , wherein the mutation-inducing dose of imatinib is a single dose. 
     
     
       11. A culture model of chronic myelogenic leukemia (CML) acquired chemoresistant cells, the model generated by:
 contacting a blast crisis CML cell line, which has previously not been exposed to imatinab before, with a single dose of imatinib sufficient to induce a BCR-ABL mutation, wherein the blast crisis CML cell line is a KCL-22 cell line; 
 maintaining a culture of the imatinib-treated cell line for a period of time until the cell line relapses and repopulates the culture; and 
 determining that the repopulated cell culture is a CML acquired chemoresistant cell line by detecting a BCR-ABL mutation. 
 
     
     
       12. The model of  claim 11 , wherein the mutation-inducing dose of imatinib is between about 2.5 μM and about 10 μM. 
     
     
       13. The model of  claim 11 , wherein the BCR-ABL mutation is a BCR-ABL kinase domain mutation. 
     
     
       14. The model of  claim 13 , wherein the BCR-ABL kinase domain mutation is a T315I BCR-ABL mutation. 
     
     
       15. The model of  claim 14 , wherein the BCR-ABL kinase domain mutation is a E255K or a Y253H BCR-ABL mutation or a combination thereof. 
     
     
       16. The model of  claim 11 , wherein the BCR-ABL kinase domain mutation is a E255K or a Y253H BCR-ABL mutation or a combination thereof. 
     
     
       17. The model of  claim 11 , wherein the BCR-ABL mutation is stable. 
     
     
       18. The model of  claim 11 , wherein the period of time is about two to three weeks.